WO2019115576A1 - Methods of treating heart failure - Google Patents

Abstract

Worldwide, 1% to 2% of the general adult population have heart failure (HF), which is accompanied by reduced quality of life, high morbidity, mortality, and significant financial costs. Recently, the potential role of the gut in the pathophysiology of heart failure (HF) has recently begun to attract increased attention. In this context, the inventors demonstrated that treatment with Lactobbacillus reuteri ameliorates mice survival following myocardial infarction and that said treatment prevented the decrease of both cardiac function, (estimated by the % of ejection fraction) and cardiac remodeling. Moreover, the inventors show that ligands of aryl hydrocarbon receptor (AHR) that comprise some metabolites produced by Lactobacillus reuteri are also particularly suitable for the treatment of heart failure. Accordingly, the present invention relates to methods of treating heart failure in a subject in need thereof comprising administering to the subject an effective amount of Lactobacillus reuteri or at least one ligand of AHR.

Description

METHODS OF TREATING HEART FAILURE

FIELD OF THE INVENTION:

The present invention relates to methods of treating heart failure.

BACKGROUND OF THE INVENTION:

Worldwide, 1% to 2% of the general adult population have heart failure (HF), which is accompanied by reduced quality of life, high morbidity, mortality, and significant financial costs. Existing therapies such as renin angiotensin aldosterone system (RAAS) inhibition, b- blockade, and angiotensin receptor-neprilysin inhibitors reduce hospitalization and mortality risk in patients with HF. Despite important cardiovascular benefits with the use of these agents, patients still have an increased risk for morbidity and mortality. Identification of novel therapeutic strategies to improve symptoms, reduce mortality, recurrent hospitalization, and acute decompensation is therefore critical to advance outcomes in patients with HF. The gut microbiota has emerged as a central factor affecting human health and disease, and cardiovascular diseases are no exception. The potential role of the gut in the pathophysiology of heart failure (HF) has recently begun to attract increased attention. For example, a recent study used 16S ribosomal RNA gene sequencing of fecal samples from HF patients to determine that gut microbial dysbiosis is associated with HF (Kamo T, Akazawa H, Suda W, et al. Dysbiosis and compositional alterations with aging in the gut microbiota of patients with heart failure. PFoS One. 20l7;l2:e0l74099). Importantly, this result is suggestive of the potential impact that the gut microbiota may have on the pathophysiological processes involved in HF. In addition, a wide variety of metabolites derived from gut microbes may also influence HF.

SUMMARY OF THE INVENTION:

The present invention relates to methods of treating heart failure. In particular, the present invention is defined by the claims.

DETAILED DESCRIPTION OF THE INVENTION:

The first object of the present invention relates to a method of treating heart failure in a subject in need thereof comprising administering to the subject an effective amount of Lactobacillus reuteri.

As used herein, the terms "subject," and "patient," used interchangeably herein, refer to a mammal, particularly a human who has been previously diagnosed with heart failure or who is at risk for having or developing heart failure. Typically, the subject suffers from a cardiovascular disease leading to heart failure. In some embodiments, the subject has experienced a myocardial infarction, in particular an acute myocardial infarction.

As used herein, the term "heart failure" or“HF has its general meaning in the art and embraces congestive heart failure and/or chronic heart failure. Functional classification of heart failure is generally done by the New York Heart Association Functional Classification (Criteria Committee, New York Heart Association. Diseases of the heart and blood vessels. Nomenclature and criteria for diagnosis, 6th ed. Boston: Little, Brown and co, 1964; 114). This classification stages the severity of heart failure into 4 classes (I -IV). The classes (I -IV) are: Class I: no limitation is experienced in any activities; there are no symptoms from ordinary activities; Class II: slight, mild limitation of activity; the patient is comfortable at rest or with mild exertion;Class III: marked limitation of any activity; the patient is comfortable only at rest; Class IV : any physical activity brings on discomfort and symptoms occur at rest.

In some embodiments, the method of the invention may reduce death or hospitalization in subjects by treating the heart failure. Moreover, the method of the invention will achieve improved cardiac condition of post- myocardial infarction (MI) subjects. In particular the method of the invention is suitable for reducing the risk or progression of heart failure, increasing the left ventricle ejection fraction (LVEF), inhibiting left ventricle enlargement, reducing left ventricle end systolic volume, reducing left ventricle end diastolic volume, ameliorating left ventricle dysfunction, improving myocardial contractibility.

As used herein, the term "treatment" or "treat" refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By "therapeutic regimen" is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., pain, disease manifestation, etc.]).

As used herein, the term“ Lacotbacillus reuteri” or“L. reuteri” has its general meaning in the art and refers to a Gram-positive bacterium that naturally inhabits the gut of mammals and that belongs to the Lactobacillus genus. Any strain of L. reuteri may be used according to the invention. In some embodiments, the L. reuteri is Lactobacillus reuteri DSM 17938, the L. reuteri strain owned by Biogaia AB, Sweden, having the scientific strain designation DSM 17938, formerly L reuteri ATCC 55730. The DSM identification refers to the DSMZ Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Inhoffenstr. 7b, D-38124 Braunschweig, Germany. DSM 17938. Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Inhoffenstr. 7b D-38124 Braunschweig - Germany. Other examples of L. reuteri suitable for use according to the invention L. reuteri are ATCC PTA 6475, L. reuteri ATCC PTA 4659 and L. reuteri ATCC PTA 5289 (available from Biogaia, Sweden), L. reuteri RC-14 (sold by Christian Hansen, France), L. reuteri NCIMB 30242 (sold as supplement called Cardioviva , by Micropharma Ltd., Canada) and L. reuteri DSMZ 17648 (sold under the commercial name Pylopass, by Lonza, Switzerland). In some embodiments, the L. reuteri is the Lactobacillus reuteri strain 93a that has been deposited under the Budapest Treaty at DSMZ (Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany) on December 11, 2015, and has been given the accession number DSM 32229. In some embodiments, the L. reuteri is the Lactobacillus reuteri strain F33 that has been deposited under the Budapest Treaty at DSMZ (Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany) on December 11, 2015, and has been given the accession number DSM 32232. In some embodiments, the L. reuteri is the Lactobacillus reuteri strain C30 has been deposited under the Budapest Treaty at DSMZ (Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany) on December 11, 2015, and has been given the accession number DSM 32230. In some embodiments, the L. reuteri is the Lactobacillus reuteri strain D276 that has been deposited under the Budapest Treaty at DSMZ (Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures, Inhoffenstr. 7B, D-38124 Braunschweig, Germany) on December 11, 2015, and has been given the accession number DSM 32231.

In some embodiments, the Lactobacillus reuteri is a probiotic strain. As used herein the term“probiotic” is meant to designate live microorganisms which, they are integrated in a sufficient amount, exert a positive effect on health, comfort and wellness beyond traditional nutritional effects. Probiotic microorganisms have been defined as“Live microorganisms which when administered in adequate amounts confer a health benefit on the host” (FAO/WHO 2001). As used herein the expression“probiotic Lactobacillus reuteri strain” denotes a Lactobacillus reuteri strain that has a beneficial effect on the health and well-being of the host.

In some embodiments, the probiotic Lactobacillus reuteri is a viable probiotic Lactobacillus reuteri strain. The expression“viable probiotic Lactobacillus reuteri strain” means a microorganism which is metabolically active and that is able to colonize the gastro- intestinal tract of the subject.

In some embodiments, the probiotic Lactobacillus reuteri strain of the present invention is a non-viable probiotic Lactobacillus reuteri strain consisting of a mixture of bacterial fragments. In some embodiments, the mixture of bacterial fragments of the present invention consists of proteins from the Lactobacillus reuteri strain.

In some embodiments, the probiotic Lactobacillus reuteri strain of the present invention is selected from food grade bacteria. "Food grade bacteria" means bacteria that are used and generally regarded as safe for use in food.

Typically, the Lactobacillus reuteri strain of the present invention is produced with any appropriate culture medium well known in the art. Various fermentation media are suitable according to the invention, such as (but not limited to) e.g. firstly an industrial medium, in which the strain(s) is/are grown, and that is used as is or after concentration (e.g. drying) or after addition to another food base or product. Alternatively, bacterial cells, or bacterial cells with medium (e.g. the fermentation broth), or fractions of such cell comprising medium (i.e. medium with said bacterial strain/s) may be used. The cells or the cell comprising medium comprise live or viable bacterial cells and/or dead or non-viable bacterial cells of the strain(s). The medium may thus be treated by, but not limited to, heating or sonication. Also lyophilized, or frozen, bacteria and/or cell-free media (which may be concentrated) are encompassed in the methods for preparing the Lactobacillus reuteri strain of the present invention. In some embodiments, the Lactobacillus reuteri strain of the present invention is encapsulated in order to be protected against the stomach. Accordingly, in some embodiments the Lactobacillus reuteri strain of the present invention is formulated in compositions in an encapsulated form so as significantly to improve their survival time. In such a case, the presence of a capsule may in particular delay or prevent the degradation of the microorganism in the gastrointestinal tract. It will be appreciated that the compositions of the present embodiments can be encapsulated into an enterically-coated, time-released capsule or tablet. The enteric coating allows the capsule/tablet to remain intact (i.e., undissolved) as it passes through the gastrointestinal tract, until such time as it reaches the intestine. Methods of encapsulating live bacterial cells are well known in the art (see, e.g., U.S. patents to General Mills Inc. such as U.S. Pat. No. 6,723,358). For example, micro-encapsulation with alginate and Hi-Maize™ starch followed by freeze-drying has been proved successful in prolonging shelf-life of bacterial cells in dairy products [see, e.g., Kailasapathy et al. Curr Issues Intest Microbiol. 2002 September; 3(2):39-48] Alternatively encapsulation can be done with glucomannane fibers such as those extracted from Amorphophallus konjac. Alternatively, entrapment of viable probiotic in sesame oil emulsions may also be used [see, e.g., Hou et al. J. Dairy Sci. 86:424- 428] In some embodiments, agents for enteric coatings are preferably methacrylic acid- alkyl acrylate copolymers, such as Eudragit® polymers. Poly(meth)acrylates have proven particularly suitable as coating materials. EUDRAGIT® is the trade name for copolymers derived from esters of acrylic and methacrylic acid, whose properties are determined by functional groups. The individual EUDRAGIT® grades differ in their proportion of neutral, alkaline or acid groups and thus in terms of physicochemical properties. The skillful use and combination of different EUDRAGIT® polymers offers ideal solutions for controlled drug release in various pharmaceutical and technical applications. EUDRAGIT® provides functional films for sustained-release tablet and pellet coatings. The polymers are described in international pharmacopeias such as Ph.Eur., USP/NF, DMF and JPE. EUDRAGIT® polymers can provide the following possibilities for controlled drug release: gastrointestinal tract targeting (gastroresistance, release in the colon), protective coatings (taste and odor masking, protection against moisture) and delayed drug release (sustained-release formulations). EUDRAGIT® polymers are available in a wide range of different concentrations and physical forms, including aqueous solutions, aqueous dispersion, organic solutions, and solid substances. The pharmaceutical properties of EUDRAGIT® polymers are determined by the chemical properties of their functional groups. Poly(meth)acrylates, soluble in digestive fluids (by salt formation) EUDRAGIT® L (Methacrylic acid copolymer), S (Methacrylic acid copolymer), FS and E (basic butylated methacrylate copolymer) polymers with acidic or alkaline groups enable pH-dependent release of the active ingredient. Applications: from simple taste masking via resistance solely to gastric fluid, to controlled drug release in all sections of the intestine. Poly(meth)acrylates, insoluble in digestive fluids: EUDRAGIT® RL and RS (ammonio methacrylate copolymers) polymers with alkaline and EUDRAGIT® NE polymers with neutral groups enable controlled time release of the active by pH-independent swelling. Enteric EUDRAGIT® coatings provide protection against drug release in the stomach and enable controlled release in the intestine. The dominant criterion for release is the pH-dependent dissolution of the coating, which takes place in a certain section of the intestine (pH 5 to over 7) rather than in the stomach (pH 1-5). For these applications, anionic EUDRAGIT® grades containing carboxyl groups can be mixed with each other. This makes it possible to finely adjust the dissolution pH, and thus to define the drug release site in the intestine. EUDRAGIT® L and S grades are suitable for enteric coatings. EUDRAGIT® FS 30 D (aqueous dispersion of an anionic copolymer based on methyl acrylate, methyl methacrylate and methacrylic acid) is specifically used for controlled release in the colon.

In some embodiments, the Lactobacillus reuteri strain of the present invention is administered to the subject in the form of a food composition. In some embodiments, the food composition is selected from complete food compositions, food supplements, nutraceutical compositions, and the like. The composition of the present invention may be used as a food ingredient and/or feed ingredient. The food ingredient may be in the form of a solution or as a solid— depending on the use and/or the mode of application and/or the mode of administration.

As used herein, the term“food” refers to liquid (i.e. drink), solid or semi-solid dietetic compositions, especially total food compositions (food-replacement), which do not require additional nutrient intake or food supplement compositions. Food supplement compositions do not completely replace nutrient intake by other means. Food and food supplement compositions are for example fermented dairy products or dairy-based products, which are preferably administered or ingested orally one or more times daily. Fermented dairy products can be made directly using the bacteria according to the invention in the production process, e.g. by addition to the food base, using methods known per se. In such methods, the strain(s) of the invention may be used in addition to the micro-organism usually used, and/or may replace one or more or part of the micro-organism usually used. For example, in the preparation of fermented dairy products such as yoghurt or yoghurt-based drinks, a bacterium of the invention may be added to or used as part of a starter culture or may be suitably added during such a fermentation. Optionally the bacteria may be inactivated or killed later in the production process. Fermented dairy products include milk-based products, such as (but not limited to) deserts, yoghurt, yoghurt drinks, quark, kefir, fermented milk-based drinks, buttermilk, cheeses, dressings, low fat spreads, fresh cheese, soy-based drinks, ice cream, etc. Alternatively, food and/or food supplement compositions may be non-dairy or dairy non fermented products (e.g. strains or cell-free medium in non-fermented milk or in another food medium). In some embodiments, the Lactobacillus reuteri strain of the present invention is encapsulated and dispersed in a food (e.g. in milk) or non-food medium. Non-fermented dairy products may include ice cream, nutritional bars and dressings, and the like. Non-dairy products may include powdered beverages and nutritional bars, and the like. The products may be made using known methods, such as adding an effective amount of the strain(s) and/or cell-free culture medium to a food base, such as skimmed milk or milk or a milk-based composition and fermentation as known. Other food bases to which the (compositions comprising the) bacterial cells and/or cell-free culture medium may be added are meat, meat replacers or plant bases. The composition that comprises the Lactobacillus reuteri strain of the present invention may be solid, semi-solid or liquid. It may be in the form of a food product or food supplement, e.g. in the form of tablets, gels, powders, capsules, drinks, bars, etc. For example the composition may be in the form of a powder packed in a sachet which can be dissolved in water, fruit juice, milk or another beverage.

As used herein the term“food ingredient” or“feed ingredient” includes a formulation which is or can be added to functional foods or foodstuffs as a nutritional supplement. By “nutritional food” or“nutraceutical” or“functional” food, is meant a foodstuff which contains ingredients having beneficial effects for health or capable of improving physiological functions. By“food supplement”, is meant a foodstuff having the purpose of completing normal food diet. A food supplement is a concentrated source of nutrients or other substances having a nutritional or physiological effect, when they are taken alone or as a combination in small amounts. According to the invention, “functional food” summarizes foodstuff and corresponding products lately developed to which importance is attributed not only due to them being valuable as to nutrition and taste but due to particular ingredient substances. According to the invention, the middle- or long-term maintenance and promotion of health are of importance. In this context, non-therapeutic uses are preferred. The terms“nutriceuticals”,“foodsceuticals” and“designer foods”, which also represent embodiments of the invention, are used as synonyms, partly, however, also in a differentiated way. The preventive aspect and the promotion of health as well as the food character of the products are, however, best made clear by the term functional food. In many cases, these relate to products accumulated by assortment and selection (as is also the case in the present invention), purification, concentration, increasingly also by addition. Isolated effective substances, in particular in form of tablets or pills, are not included. Although there is no legal definition of a functional food, most of the parties with an interest in this area agree that they are foods marketed as having specific health effects beyond basic nutritional effects. Accordingly, functional foods are ordinary foods that have components or ingredients (such as those described herein) incorporated into them that impart to the food a specific functional e.g. medical or physiological benefit other than a purely nutritional effect.

In some embodiments, the drink is a functional drink or a therapeutic drink, a thirst- quencher or an ordinary drink. By way of example, the composition of the present invention can be used as an ingredient to soft drinks, a fruit juice or a beverage comprising whey protein, health teas, cocoa drinks, milk drinks and lactic acid bacteria drinks, yoghurt and drinking yoghurt, cheese, ice cream, water ices and desserts, confectionery, biscuits cakes and cake mixes, snack foods, balanced foods and drinks, fruit fillings, care glaze, chocolate bakery filling, cheese cake flavoured filling, fruit flavoured cake filling, cake and doughnut icing, instant bakery filling creams, fillings for cookies, ready-to-use bakery filling, reduced calorie filling, adult nutritional beverage, acidified soy/juice beverage, aseptic/retorted chocolate drink, bar mixes, beverage powders, calcium fortified soy/plain and chocolate milk, calcium fortified coffee beverage.

In some embodiments, the composition that comprises the Lactobacillus reuteri strain of the present invention is used with yoghurt production, such as fermented yoghurt drink, yoghurt, drinking yoghurt, cheese, fermented cream, milk based desserts and others. Suitably, the composition can be further used as an ingredient in one or more of cheese applications, meat applications, or applications comprising protective cultures.

The food composition that comprises the Lactobacillus reuteri strain of the present invention typically comprises carriers or vehicles.“Carriers” or“vehicles” mean materials suitable for administration and include any such material known in the art such as, for example, any liquid, gel, solvent, liquid diluent, solubilizer, or the like, which is non-toxic and which does not interact with any components of the composition in a deleterious manner. Examples of nutritionally acceptable carriers include, for example, water, salt solutions, alcohol, silicone, waxes, petroleum jelly, vegetable oils, polyethylene glycols, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, petroethral fatty acid esters, hydroxymethyl-cellulose, polyvinylpyrrolidone, and the like. In some embodiments, the food composition that comprises the Lactobacillus reuteri strain of the present invention comprises an amount of dietary fibres. Dietary fibre passes through the small intestine undigested by enzymes and functions as a natural bulking agent and laxative. Dietary fibre may be soluble or insoluble and in general a blend of the two types is preferred. Suitable sources of dietary fibre include soy, pea, oat, pectin, guar gum, gum Arabic, ffuctooligosaccharides, galacto-oligosaccharides, sialyl-lactose and oligosaccharides derived from animal milks. In some embodiments, the dietary fiber is selected among mannans. Mannans (such as glucomannans and galactomannans), such as guar gum, locust bean gum, konjac, and xanthan gum, are present in some plant cell walls. The glucomannans are generally comprised of ( 1 -4)-b-1 inked glucose and mannose units, while the galactomannans are generally comprised of a (l-4)-P-mannan backbone substituted with single units of (l-6)-a- galactose. Many endospermic legumes, such as guar and locust bean, contain galactomannans in the endosperm during seed development. Glucomannans have also been found as a minor component of cereal grains.

In some embodiments, the composition that comprises the Lactobacillus reuteri strain of the present invention contains emulsifiers. Examples of food grade emulsifiers typically include diacetyl tartaric acid esters of mono- and di-glycerides, lecithin and mono- and di- glycerides. Similarly suitable salts and stabilisers may be included.

In some embodiments, the food composition that comprises the Lactobacillus reuteri strain of the present invention contains at least one prebiotic. "Prebiotic" means food substances intended to promote the growth of the Lactobacillus reuteri strain of the present invention in the intestines. The prebiotic may be selected from the group consisting of oligosaccharides and optionally contains fructose, galactose, mannose, soy and/or inulin; and/or dietary fibers.

In some embodiments, the composition that comprises the Lactobacillus reuteri strain of the present invention contains protective hydrocolloids (such as gums, proteins, modified starches), binders, film forming agents, encapsulating agents/materials, wall/shell materials, matrix compounds, coatings, emulsifiers, surface active agents, solubilizing agents (oils, fats, waxes, lecithins etc.), adsorbents, carriers, fillers, co-compounds, dispersing agents, wetting agents, processing aids (solvents), flowing agents, taste masking agents, weighting agents, jellifying agents, gel forming agents, antioxidants and antimicrobials. The composition may also contain conventional pharmaceutical additives and adjuvants, excipients and diluents, including, but not limited to, water, gelatine of any origin, vegetable gums, ligninsulfonate, talc, sugars, starch, gum arabic, vegetable oils, polyalkylene glycols, flavouring agents, preservatives, stabilizers, emulsifying agents, buffers, lubricants, colorants, wetting agents, fillers, and the like. In all cases, such further components will be selected having regard to their suitability for the intended recipient.

In some embodiments, the administration of the Lactobacillus reuteri strain is repeated, for example, 2 to 3 times a day, for one day or more and generally for a sustained period of at least 4 days, or even 4 to 15 weeks, with, where appropriate, one or more periods of interruption. In some embodiments, the Lactobacillus reuteri strain is administered simultaneously or sequentially one meal of the subject.

As used herein, the term "effective amount" refers to a quantity sufficient of the Lactobacillus reuteri strain to achieve the beneficial effect (e.g. treating heart failure). In the context of the present invention, the amount of the Lactobacillus reuteri strain administered to the subject will depend on the characteristics of the individual, such as general health, age, sex, body weight... The skilled artisan will be able to determine appropriate dosages depending on these and other factors. For example, the Lactobacillus reuteri strain shall be able to generate a colony is sufficient to generate a beneficial effect on the subject. If the Lactobacillus reuteri strain is administered in the form of a food product, it typically may comprise between 10³ and 10¹² cfu of the Lactobacillus reuteri strain of the present invention per g of the dry weight of the food composition.

Without to be bound by any particular theory the inventors believe that L. reuteri produces some particular metabolites that are suitable for the treatment of heart failure. In particular, the inventor show that some ligands of aryl hydrocarbon receptor (AHR) are suitable for treating heart failure (see the EXAMPLE).

Accordingly, a further object of the present invention relates to a method of treating heart failure in a subject in need thereof comprising administering to the subject a therapeutically effective amount of at least one ligand of aryl hydrocarbon receptor (AHR).

As used herein, the“ayrl hydrocarbon receptor” or“AHR” has its general meaning in the art and is a ligand activated transcription factor of the basic region helix-loop-helix- PER/ARNT/SIM homology family. Accordingly, the term“ligand of AHR” refers to any compound natural or not that is capable to binding AHR and promotes activation of the signaling pathway of AHR. The prototypic signaling pathway of AHR-mediated transcriptional activity is characterized by transcription of a battery of drug-metabolizing enzymes, which includes cytochrome P450 enzymes 1A1, 1A2, and 1B1.

In some embodiments, the ligand is selected from the group consisting of 2, 3,7,8- Tetrachlorodibenzo-p-dioxin (TCDD), lndole-3-carbinol (I3C), lndole-3 -acetonitrile (I3ACN), 3,3-Diindolylmethane (DIM), 2-(Indol-3-ylmethyl)-3,3’-diindolylmethane (Ltr-l), Indolo[3,2- bjcarbazole (ICZ), 2-(l ' H-indole-3 ' -carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE), Indole, lndole-3 -acetic acid (IAA), lndole-3 -aldehyde (IAld), Tryptamine, 3-Methyl- indole (skatole), lndoxyl-3 -sulfate (I3S), Kynurenine (Kyn), Kynurenic acid (KA), Xanthurenic acid, Cinnabarinic acid (CA), and 6-Formylindolo[3,2-b]carbazole (FICZ).

Typically the ligand of AHR is administered to the subject in a food composition as described above. In some embodiments, the ligand of AHR is administered to the subject in a form of a pharmaceutical composition. For instance, the ligand of AHR may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions. "Pharmaceutically" or "pharmaceutically acceptable" refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. In the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, the active principle, alone or in combination with another active principle, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms. Typically, the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The ligand of AHR can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin. Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the typical methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents to a small tumor area. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.

FIGURES:

Figure 1 demonstrated that heart failure is associated with a decreased fecal expression of L. reuteri in mice following 3 days of myocardial infarction, as compared to sham-operated mice. *: P<0.05 versus Sham.

Figure 2 demonstrated that L. reuteri treatment ameliorates mice survival following myocardial infarction. *: P<0.05 versus MI.

Figure 3 shows the left ventricular ejection fraction (EF %) (Figure 3 A) and cardiac volumes of mice which underwent a thoracotomy but no coronary ligation (Sham) (Figure 3B), mice which underwent a thoracotomy with coronary ligation (MI d+28), mice which underwent a thoracotomy with coronary ligation and were treated with L. reuteri 7 days before and during 28 days after the surgery (MI d+28 + L-Reu d-7), as disclosed in the example below a: P<0.05 versus Sham; b: P<0.05 versus MI d+28.

Figure 4 shows the left ventricular ejection fraction (EF %) of mice which underwent a thoracotomy but no coronary ligation (Sham), mice which underwent a thoracotomy with coronary ligation (MI d+28), mice which underwent a thoracotomy with coronary ligation and were treated with Indole 7 days before and during 28 days after the surgery (MI d+28 + Indole d-7), as disclosed in the example below a: P<0.05 versus Sham; b: P<0.05 versus MI d+28.

Figure 5 shows the left ventricular ejection fraction (EF %) of mice which underwent a thoracotomy but no coronary ligation (Sham), mice which underwent a thoracotomy with coronary ligation (MI d+28), mice which underwent a thoracotomy with coronary ligation and were treated with FICZ 7 days before and during 28 days after the surgery (MI d+28 + FICZ d- 7), as disclosed in the example below a: P<0.05 versus Sham; b: P<0.05 versus MI d+28. EXAMPLE:

Methods

Treatment of mice

Five groups of mice were formed.

- The first control group of mice (Sham) underwent a thoracotomy but no coronary ligation.

- The second group of mice (MI d+28) underwent a thoracotomy with coronary ligation.

- The third group of mice (MI d+28 + L-Reu d-7) underwent a thoracotomy with coronary ligation and was treated with Lactobacillus reuteri 7 days before and during 28 days after the surgery.

- The fourth group of mice (MI d+28 + Indole d-7) underwent a thoracotomy with coronary ligation and was treated with Indole 7 days before and during 28 days after the surgery.

- The fith group of mice (MI d+28 + FICZ d-7) underwent a thoracotomy with coronary ligation and was treated with FICZ (5,1 l-Dihydroindolo[3,2-£>]carbazole-6-carboxaldehyde), a specific agonist of Ahr receptor, 7 days before and during 28 days after the surgery.

The treatment with L. reuteri was administered daily by gavage (3.9 10⁹ cfu /mice /day, ATCC).

The treatment with Indole was administered daily by gavage (400 pg/ 20 g BW / day, Sigma).

The treatment with FICZ was administered every 2 days by intraperitoneal injection (IP, 2.5 pg/ mice, Tocris).

Quantification of Lactobacillus reuteri DNA in mouse feces

Bacterial DNA in 200 mg of fecal samples from Sham and MI mice was purified using QIAamp DNA Stool Mini Kit (Qiagen) and subjected to SYBR Green qPCR using primers specific to Lactobacillus reuteri: Lactobacillus reuteri primers were used as follows: Forward: 5'-ACCGAGAACACCGCGTTATTT-3' (SEQ ID NO: l), Reverse: 5'-

C AT AACTT AAC C AAAC AAT C AAAG ATT GT CT-3 ' (SEQ ID NO:2).

Cardiac surgery

Myocardial infarction (MI) was performed on 8 week-old male C57BL/6J. In brief, male 8 week-old mice were anesthetized by an intraperitoneal (i.p.) injection of a cocktail of ketamine (100 mg/kg) and xylazine (10 mg/kg), intubated, and connected to a mouse ventilator (MiniVent, Harvard Apparatus, Holliston, MA). Permanent ligation of the left anterior descending artery was blocked using a segment of saline 9-0 prolene. The sham group (without ligation of the left anterior descending artery) was set up as the control group. All surgical procedures were performed under sterile conditions. Successful cardiac infarction was confirmed by apparent S-T segment elevation. 4 weeks post- surgery, echocardiography; subsequently, cardiac tissues from different regions were harvested for further analysis.

Echocardiography

Non-invasive ultrasound examination of the cardiovascular system was performed using a General Electric instrument equipped with a linear 8-l4-MHz transducer. The surgeon and echocardiographer were blinded to animal genotype.

Results

Figure 1 demonstrated that heart failure is associated with a decreased fecal expression of L. reuteri in mice following 3 days of myocardial infarction, as compared to sham-operated mice. Figure 2 demonstrated that L. reuteri treatment ameliorates mice survival following myocardial infarction. Figure 3 A and B demonstrated that L. reuteri treatment prevented the decrease of both cardiac function, (estimated by the % of ejection fraction) and cardiac remodeling (estimated by the left ventricular end-diastolic and end-systolic volumes) following 28 days of MI. Figure 4 demonstrated that Indole (first metabolite of tryptophan produced by

L. reuteri ) treatment prevented the decrease of cardiac function, (estimated by the % of ejection fraction) following 28 days of MI. Figure 5 demonstrated that FICZ treatment prevented the decrease of cardiac function, (estimated by the % of ejection fraction) following 28 days of MI.

REFERENCES:

Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

Claims

CLAIMS:

1. A method of treating heart failure in a subject in need thereof comprising administering to the subject an effective amount of Lactobacillus reuteri.

2. The method of claim 1 wherein the subject was previously diagnosed with heart failure or is at risk for having or developing heart failure.

3. The method of claim 1 wherein the subject has experienced a myocardial infarction, in particular an acute myocardial infarction.

4. The method of claim 1 wherein the Lactobacillus reuteri is a probiotic strain.

5. The method of claim 4 wherein the probiotic Lactobacillus reuteri strain is a viable probiotic Lactobacillus reuteri strain.

6. The method of claim 4 wherein the probiotic Lactobacillus reuteri strain is a non-viable probiotic Lactobacillus reuteri strain.

7. The method of claim 4 wherein the probiotic Lactobacillus reuteri strain is selected from food grade bacteria.

8. The method of claim 1 wherein the Lactobacillus reuteri strain is encapsulated in order to be protected against the stomach.

9. The method of claim 1 wherein the Lactobacillus reuteri strain is administered to the subject in the form of a food composition.

10. The method of claim 9 wherein the food composition further comprises an amount of dietary fibres.

11. A method of treating heart failure in a subject in need thereof comprising administering to the subject a therapeutically effective amount of at least one ligand of aryl hydrocarbon receptor (AHR).

12. The method of claim 11 wherein the ligand is selected from the group consisting of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), lndole-3-carbinol (I3C), lndole-3- acetonitrile (I3ACN), 3,3-Diindolylmethane (DIM), 2-(Indol-3-ylmethyl)-3,3’- diindolylmethane (Ltr-l), Indolo[3,2-b]carbazole (ICZ), 2-( 1 'H-indolc-3'-carbonyl)- thiazole-4-carboxylic acid methyl ester (ITE), Indole, lndole-3 -acetic acid (IAA), lndole-3 -aldehyde (IAld), Tryptamine, 3 -Methyl-indole (skatole), lndoxyl-3 -sulfate (13 S), Kynurenine (Kyn), Kynurenic acid (KA), Xanthurenic acid, Cinnabarinic acid (CA), and 6-Formylindolo[3,2-b]carbazole (FICZ).

13. The method of claim 11 wherein the ligand of AHR is administered to the subject in a form of a food composition.

14. The method of claim 11 wherein the ligand of AHR is administered to the subject in a form of a pharmaceutical composition.